• A Corrigendum to this article was published on 15 June 2011

This article has been updated

Abstract

Microtubules have pivotal roles in fundamental cellular processes and are targets of antitubulin chemotherapeutics1. Microtubule-targeted agents such as Taxol and vincristine are prescribed widely for various malignancies, including ovarian and breast adenocarcinomas, non-small-cell lung cancer, leukaemias and lymphomas1. These agents arrest cells in mitosis and subsequently induce cell death through poorly defined mechanisms2. The strategies that resistant tumour cells use to evade death induced by antitubulin agents are also unclear2. Here we show that the pro-survival protein MCL1 (ref. 3) is a crucial regulator of apoptosis triggered by antitubulin chemotherapeutics. During mitotic arrest, MCL1 protein levels decline markedly, through a post-translational mechanism, potentiating cell death. Phosphorylation of MCL1 directs its interaction with the tumour-suppressor protein FBW7, which is the substrate-binding component of a ubiquitin ligase complex. The polyubiquitylation of MCL1 then targets it for proteasomal degradation. The degradation of MCL1 was blocked in patient-derived tumour cells that lacked FBW7 or had loss-of-function mutations in FBW7, conferring resistance to antitubulin agents and promoting chemotherapeutic-induced polyploidy. Additionally, primary tumour samples were enriched for FBW7 inactivation and elevated MCL1 levels, underscoring the prominent roles of these proteins in oncogenesis. Our findings suggest that profiling the FBW7 and MCL1 status of tumours, in terms of protein levels, messenger RNA levels and genetic status, could be useful to predict the response of patients to antitubulin chemotherapeutics.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Change history

  • 09 March 2011

    Minor errors in the HTML and PDF were corrected on 09March 2011.

  • 21 November 2011

    Supplementary Fig. S21 contained a duplicated panel instead of the SB203580-treated cdc27 blot.

References

  1. 1.

    , , & Targeted anti-mitotic therapies: can we improve on tubulin agents? Nature Rev. Cancer 7, 107–117 (2007)

  2. 2.

    & Stuck in division or passing through: what happens when cells cannot satisfy the spindle assembly checkpoint. Dev. Cell 7, 637–651 (2004)

  3. 3.

    & The BCL-2 protein family: opposing activities that mediate cell death. Nature Rev. Mol. Cell Biol. 9, 47–59 (2008)

  4. 4.

    et al. Apoptosis initiated when BH3 ligands engage multiple Bcl-2 homologs, not Bax or Bak. Science 315, 856–859 (2007)

  5. 5.

    & (Un)expected roles of c-IAPs in apoptotic and NFκB signaling pathways. Cell Cycle 7, 1511–1521 (2008)

  6. 6.

    et al. A 20S complex containing CDC27 and CDC16 catalyzes the mitosis-specific conjugation of ubiquitin to cyclin B. Cell 81, 279–288 (1995)

  7. 7.

    Recognition and processing of ubiquitin–protein conjugates by the proteasome. Annu. Rev. Biochem. 78, 477–513 (2009)

  8. 8.

    & Deregulated proteolysis by the F-box proteins SKP2 and β-TrCP: tipping the scales of cancer. Nature Rev. Cancer 8, 438–449 (2008)

  9. 9.

    & FBW7 ubiquitin ligase: a tumour suppressor at the crossroads of cell division, growth and differentiation. Nature Rev. Cancer 8, 83–93 (2008)

  10. 10.

    & RING domain E3 ubiquitin ligases. Annu. Rev. Biochem. 78, 399–434 (2009)

  11. 11.

    , , & Mule/ARF-BP1, a BH3-only E3 ubiquitin ligase, catalyzes the polyubiquitination of Mcl-1 and regulates apoptosis. Cell 121, 1085–1095 (2005)

  12. 12.

    et al. The reversibility of mitotic exit in vertebrate cells. Nature 440, 954–958 (2006)

  13. 13.

    , , & Evidence that mitotic exit is a better cancer therapeutic target than spindle assembly. Cancer Cell 16, 347–358 (2009)

  14. 14.

    , , , & MCL1 is phosphorylated in the PEST region and stabilized upon ERK activation in viable cells, and at additional sites with cytotoxic okadaic acid or Taxol. Oncogene 23, 5301–5315 (2004)

  15. 15.

    & Cancer cells display profound intra- and interline variation following prolonged exposure to antimitotic drugs. Cancer Cell 14, 111–122 (2008)

  16. 16.

    , , , & Fbw7 regulates the activity of endoreduplication mediators and the p53 pathway to prevent drug-induced polyploidy. Oncogene 27, 4411–4421 (2008)

  17. 17.

    , & Cyclin-dependent kinase 1-mediated Bcl-xL/Bcl-2 phosphorylation acts as a functional link coupling mitotic arrest and apoptosis. Mol. Cell. Biol. 30, 640–656 (2010)

  18. 18.

    , , , & Multidrug resistance gene-1 (Pgp) expression in epithelial ovarian malignancies. Eur. J. Gynaecol. Oncol. 23, 337–340 (2002)

  19. 19.

    et al. No significant role for β tubulin mutations and mismatch repair defects in ovarian cancer resistance to paclitaxel/cisplatin. BMC Cancer 5, 101 (2005)

  20. 20.

    et al. Differential targeting of prosurvival Bcl-2 proteins by their BH3-only ligands allows complementary apoptotic function. Mol. Cell 17, 393–403 (2005)

Download references

Acknowledgements

We thank P. Ekert for FDM cell lines, J. Stinson for sequencing assistance, S. Johnson and C. Santos for assistance with obtaining patient samples, P. Haverty for bioinformatics analysis, C. Grimaldi for cloning assistance, J. Dynek for TaqMan advice, D. French for tumour analysis, C. Quan and J. Tom for peptide synthesis, I. Zilberleyb and the Baculovirus Expression Group for cloning and protein production, S. Charuvu for generating MCL1 point mutants, A. Bruce for graphics assistance, the Genentech Cancer Genome Project Team, Z. Modrusan, R. Soriano and the microarray lab for ovarian tumour data sets, K. Newton for editorial assistance, W. Wei for sharing unpublished results, and A. Eldridge, D. Kirkpatrick, D. Vucic, E. Varfolomeev, T. Goncharov, A. Cochran, O. Huang, A. Huang, Y. Pereg, A. Loktev, D. Phillips, J. Wu, M. van Delft, D. Eaton, E. Shaulian, T. Hunter, S. Cory, J. Adams, A. Strasser, R. Deshaies and G. Evan for discussions. Work in the Huang laboratory is supported by the National Health and Medical Research Council (program grant #461221, IRIISS grant #361646 and a fellowship to D.C.S.H.), the Leukemia and Lymphoma Society (SCOR 7413), the National Institutes of Health (grants CA043540 and CA80188), the Australian Cancer Research Foundation, and an Australian Research Council Australian Postdoctoral fellowship to T.O. We apologize to our colleagues whose primary work could not be cited owing to space constraints.

Author information

Author notes

    • Cynthia Lam
    •  & Toru Okamoto

    These authors contributed equally to this work.

Affiliations

  1. Department of Early Discovery Biochemistry, Genentech, South San Francisco, California 94080, USA

    • Ingrid E. Wertz
    • , Saritha Kusam
    • , Cynthia Lam
    • , Elizabeth Helgason
    • , James A. Ernst
    • , Wayne J. Fairbrother
    •  & Erin C. Dueber
  2. The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3052, Australia

    • Toru Okamoto
    •  & David C. S. Huang
  3. Department of Protein Chemistry, Genentech, South San Francisco, California 94080, USA

    • Wendy Sandoval
    • , James A. Ernst
    •  & Jennie R. Lill
  4. Department of Research Oncology, Genentech, South San Francisco, California 94080, USA

    • Daniel J. Anderson
    • , Mike Eby
    • , Lisa D. Belmont
    • , Peter K. Jackson
    • , Mary J. C. Ludlam
    • , Kevin G. Leong
    •  & Heather Maecker
  5. Department of Bioinformatics, Genentech, South San Francisco, California 94080, USA

    • Jinfeng Liu
    •  & Joshua S. Kaminker
  6. Department of Physiological Chemistry, Genentech, South San Francisco, California 94080, USA

    • Karen M. O’Rourke
    •  & Vishva M. Dixit
  7. Department of Molecular Biology, Genentech, South San Francisco, California 94080, USA

    • Kanan Pujara
    •  & Somasekar Seshagiri
  8. Department of Biochemical Pharmacology, Genentech, South San Francisco, California 94080, USA

    • Pawan Bir Kohli
    •  & Adam R. Johnson
  9. Department of Structural Biology, Abbott Laboratories, Abbott Park, Illinois 60064, USA

    • Mark L. Chiu
  10. Department of Medical Biology, University of Melbourne, Parkville, Victoria 3010, Australia

    • David C. S. Huang

Authors

  1. Search for Ingrid E. Wertz in:

  2. Search for Saritha Kusam in:

  3. Search for Cynthia Lam in:

  4. Search for Toru Okamoto in:

  5. Search for Wendy Sandoval in:

  6. Search for Daniel J. Anderson in:

  7. Search for Elizabeth Helgason in:

  8. Search for James A. Ernst in:

  9. Search for Mike Eby in:

  10. Search for Jinfeng Liu in:

  11. Search for Lisa D. Belmont in:

  12. Search for Joshua S. Kaminker in:

  13. Search for Karen M. O’Rourke in:

  14. Search for Kanan Pujara in:

  15. Search for Pawan Bir Kohli in:

  16. Search for Adam R. Johnson in:

  17. Search for Mark L. Chiu in:

  18. Search for Jennie R. Lill in:

  19. Search for Peter K. Jackson in:

  20. Search for Wayne J. Fairbrother in:

  21. Search for Somasekar Seshagiri in:

  22. Search for Mary J. C. Ludlam in:

  23. Search for Kevin G. Leong in:

  24. Search for Erin C. Dueber in:

  25. Search for Heather Maecker in:

  26. Search for David C. S. Huang in:

  27. Search for Vishva M. Dixit in:

Contributions

I.E.W., S.K., T.O., J.A.E., P.B.K., A.R.J., C.L., E.C.D., E.H., H.M. and K.G.L. designed and performed in vitro, cell-based and in vivo experiments. D.J.A. and M.J.C.L. designed and performed microscopy experiments. S.K., C.L., K.M.O., M.L.C. and M.E. made constructs. J.L. and J.K. performed bioinformatics analysis, K.P. and S.S. provided sequencing analysis. W.S. and J.R.L. designed and performed mass spectrometry experiments. I.E.W., S.K., C.L., T.O., W.S., D.J.A., M.J.C.L., K.G.L., E.C.D., H.M. and V.M.D. prepared the manuscript and figures. W.S., L.D.B., P.K.J., W.J.F., D.J.A., P.B.K., A.R.J., M.J.C.L., H.M., D.C.S.H. and I.E.W. contributed to the study design and data analysis.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Ingrid E. Wertz.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    The file contains Supplementary Figures 1-39 with legends, Supplementary Methods, additional references and Supplementary Tables 1-2.

  2. 2.

    Supplementary Figure

    This file contains a corrected version of Supplementary Figure S21 and legend. This file was uploaded on 21 November 2011.

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nature09779

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.